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8 thoughts on “C70 – Crosbie-Villaseca”
I thought you did a really nice job here, my question is what role you predict Socax to have in DNA repair?
I personally have no clue based off of my results since this experiment just shows that Socax is upregulated in the presence of DNA damage. The results do call for more experimentation, though, to find the exact role Socax may have in DNA repair. Using CRISPR to knock out Socax would be an effective way of doing that.
Hi Alex! Great presentation, what caused the unexpected results in the controls?
What most likely caused the controls to yield unexpected results was a miscalculation or misloading of the amount of cDNA in those wells. With more cDNA there are more strands for primers to anneal to, which would simply result in more expression (brighter band) in the gel. Had I loaded the same amount of cDNA in each well I believe we would see a brighter band in the Rad51 + HU well and the same brightness bands for both Ftt18 wells.
Great job. Was there any specific reason for choosing socax
Socax shares a very similar gene expression profile with Rad51, a protein that we know is involved in DNA repair. This doesn’t necessarily mean that Socax is directly involved in DNA repair but could be involved in mitosis, which proteins involved in that process can share similar gene expression profiles as well. Since this lab is centered around DNA damage, we tested Socax to find results that may suggest a role in DNA repair.
Hi, great presentation! How did you decide on using the positive controls that you did?
Ftt18 is a great positive control since it amplifies the RPP0 gene which has an intron in between its primers. For the primer validation it helped to show that in the presence and absence of an intron, there would be two different fragment sizes which holds as a reference for the Socax gene + primers. If Socax showed two different band sizes it would look somewhat similar to that of the Ftt18 wells. For the DNA damage part of the experiment, we used Ftt18 again as a positive control to show that primers are annealing to the DNA, and Rad51 as a positive control to show what greater expression looks like after DNA damage is induced compared to what it would look like without DNA damage. They just serve as good references that the gel is loaded properly and helps visualize the results of Socax.